Pharmaceutical composition based on a hepatoprotector and a prebiotic, and production and application thereof

ABSTRACT

The invention relates to medicine, hepatology and pharmacology and can be used for producing and using a pharmaceutical composition based on a hepatoprotector and a prebiotic for treating and preventing liver diseases which are caused by lipid-cholesterol exchange and selected from the following group: cholelithiasis mainly with cholesterol stones, alcoholic and non-alcoholic steatohepatitis, biliary cirrhosis, cholesterol imbibition gallbladder and drug-induced and toxic liver damage. The pharmaceutical composition is administered by mouth and contains a hepatoprotector and a prebiotic taken, as an active agent, in therapeutically effective doses. The invention contributes to the liver&#39;s functional recovery in a short time and prevents disease recidivation owing to the recovery of cholesterol exchange and intestinal biocenosis as a result of the synergistic interaction of a hepatoprotector and a prebiotic, thereby also preventing hepatoprotector side effects.

CROSS-REFERENCE TO RELATED APPLICATIONS

The present patent application is a continuation of the U.S. patentapplication Ser. No. 12/921,383 filed Sep. 7, 2010, which is Nationalstage application of PCT application PCT/RU2008/000122 filed Mar. 4,2008.

FIELD OF INVENTION

The invention relates to medicine, particularly to hepatology andpharmacology. It can be used for production and application of apharmaceutical composition based on a hepatoprotector and a prebioticfor treatment and prevention of liver diseases selected from thefollowing group: cholelithiasis, fatty hepatosis and nonalcoholicsteatohepatitis, primary biliary cirrhosis, gallbladder cholesterosis,and drug-induced and toxic liver injury.

BACKGROUND OF THE INVENTION

The urgency of the issue is due to higher frequency and severity ofdiseases of liver, the main organ for detoxification of exogenoustoxins. The increased incidence of liver diseases is caused byenvironmental troubles in most world regions.

An additional cause which is directly related to the effect ofenvironmental factors is reduced immunity of the population, resultingin significant increase of infectious liver lesions, and first of all ofviral hepatitises.

In the case of viral hepatitis, the source of infection is a sickperson, and the infection transmission route is either fecal-oral orparenteral, depending on the type of virus-A, B, C, D, G, or E.Population's susceptibility to this infection is high.

Regardless of the sight of entry, the virus eventually gets in the liverwhere it has direct toxic effect on liver cells, combined withimmune-mediated damage of cell membranes. In all forms of viralhepatitis, a serious frequent complication is disturbance of normalprocesses of formation and flow of bile, the so-called cholestaticsyndrome accompanied by jaundice. It is most often manifested in thecase of viral hepatitis A (VHA)—“enteric” viral hepatitis and enterichepatitis E, wherein the frequency of jaundiced forms is 100%.

In severe forms of acute viral hepatitises (AVH) flow and exacerbationsof chronic hepatisises, the disturbance of structure and functionalactivity of biliary ducts is one of the reasons of development of asevere complication-biliary cirrhosis.

In addition to viral hepatitises, a large share of liver diseases is dueto the effect of food toxicants (alcohol, other toxic substances, andvarious medicinal agents).

One of the earliest pathological complications in toxic liver injury issteatohepatitis—the result of disturbance of the normal balance betweenthe input of fats in the body and their metabolism.

It should be emphasized that disturbance of normal processes offormation and biliary passage of bile is one of widespread consequencesof the effect of high doses of a number of drugs (antibiotics,sulfanilamides, chlorpromazine, histamine receptor and estrogenblockers, and cytostatics).

During the last decade, a so-called autoimmune hepatitis, the result ofdeep disturbance in the cell immunity system, is being diagnosed moreand more often. Its most severe consequence is primary biliarycirrhosis.

Disturbance of bile production and excretion processes is most vividlypronounced in the form of cholelithiasis, wherein excessive accumulation(congestion) of bile in the gallbladder with subsequent formation ofconcrements (choleliths or gallstones) is observed.

In all of the above liver diseases, an important ethiologic andpathogenic factor is disturbance of normal processes of bile acids (BA)metabolism, one of the most important factors of normal digestion.

BAs are formed in liver from cholesterol (Hofmarm A F. Bile acidsecretion, bile flow and biliary lipid secretion in humans. Hepatologi.1990; 12; 17S; Meier P J. The bile salt secretory polarity ofhepatocytes. J. Hepatol. 1989; 9: 124).

Main BAs detected in human bile are cholic acid (CA) (3a, 7a,12a-trioxy-5b-cholanic acid), chenodeoxycholic acid (CDCA), deoxycholicacid (DCA) (3a, 12a-dioxy-5b-cholanic acid). Stereoisomeres of cholicand deoxycholic acids in the form of allocholic (ACA), ursodeoxycholic(UDCA) and lithocholic (LCA) acids have been detected in bile inconsiderably smaller quantities.

CA and CDCA, the so-called primary BAs, form in the liver duringoxidation of cholesterol, and DCA and LCA form in the intestine fromprimary BAs due to the effect of enzymes of intestinal floramicroorganisms.

Normal quantitative ratio of CA, CDCA and DCA in bile is 1:1:0.6.

In bladder bile BAs are mainly present in the form of binarycompounds-conjugates. In the intestine, mainly in the ileum, BAs areabsorbed into blood, return with blood to the liver and are againsecreted within bile—this is the so-called portal-biliar circulation ofBAs; therefore, 85-90% of the entire amount of Bas contained in bile areBAs absorbed in the intestine.

Portal-biliar circulation of BAs facilitates easy absorption of BAconjugates in the intestine, because they are water-soluble; in theprocess, 10-15% of the total amount of BA break down in the intestinedue to the effect of enzymes of intestine flora microorganisms, and theproducts of their degradation are excreted with stool.

By emulsifying fats, BAs thus ensure absorption of insoluble fatty acidsand cholesterol in the small intestine, as well as of vitamins B, K, Eand calcium salts.

In addition, BAs have strong choleretic effect, stimulate intestinalmotility, and also have bacteriostatic and anti-inflammatory effect.Taking the above into account, a possible component of the method fortreatment and prevention of a number of pathologic conditions of theliver is the use of bile acid preparations, and first of all UDCA.

UDCA is a tertiary bile acid; it was first found in Chinese bear bile in1902. UDCA has been used in medicine for several centuries. As long agoas in ancient China, dried bear bile was prescribed for treatment ofstomach, intestine and liver diseases. UDCA is formed, due to the actionof bacterial enzymes, from 7-keto-litocholic acid that enters the liverfrom the small intestine.

Herein, all chemical formulae of UDCA and hydrophobic CDCA are identical(C24H4O4).

Using UDCA for treatment of, among others, liver diseases results in adose-dependent change of the above ratio of bile acids: UDCA becomes themain bile component whereas the content of CDCA and other BAs decreases.Lower accumulation of UDCA in bile is observed in patients with liverdiseases, which may be related to reduced absorption due to reducedformation of endogenous micellae from bile acids in duodenal bile or toreduced secretion of bile acids themselves.

As has been stated earlier, UDCA and LCA are detected in human bile invery insignificant amounts (0.1%-5%).

Despite good absorption of UDCA in the intestine, its level in bloodplasma remains relatively low due to fast liver clearance, becauseeffective conjugation of UDCA with glycine, taurine, N-acetoglucosaine,glucuronic acid and sulfate takes place in the liver.

The effect of UDCA on cholesterol in bile is a complex one: it reducescholesterol absorption in the intestine, its synthesis in the liver andsecretion into bile. However, there is no noticeable decrease ofcholesterol level in blood due to the effect of UDCA.

UDCA and its conjugates that have not been absorbed in the smallintestine are metabolized by indigenous bacteria in the small intestinedistal area and in the colon.

In the intestine, UDCA is broken down and dehydroxylated intolithocholic acid (LCA). LCA, whose content in human blood is very low,is formed in the small intestine due to the action of microflora duringthe process of utilization of numerous fats; from the small intestineLCA enters the colon and rectum, where it is partially absorbed, andenters the liver.

In the liver LCA bonds with sulfate anions and then with glycine andtaurine, and this way is released in bile. Its derivatives are littleabsorbed in the intestine and excreted with stool.

Such process is an efficient mechanism for elimination of toxic LCA fromthe body.

CDCA determines decrease of activity of A-oxyreductase3-hydroxy-3-methylglutarylcoenzyme—an enzyme that participates in thesynthesis of cholesterol; it also facilitates decrease of cholesterolabsorption in the intestine, which results in changing the ratio of bileacids and cholesterol towards prevailing of CDCA bile acids in thecommon pool.

The above mechanism predetermines the use of CDCA when dissolvinggallstones that mainly consist of cholesterol.

Deoxycholic acid (DCA) is a bile acid that is formed in person'sintestine due to action of intestine microflora enzymes, absorbed intoblood and secreted by the liver with bile. It is assumed thathydrophobic DCA salt can be the link between disturbed intestinalmotility and bile lithogenicity. The main bile acids in humans are CAand CDCA-primary bile acids synthesized in the liver from cholesterol.

Secondary DCA is formed from cholic acid in the small intestine distalareas and in the colon due to action of intestine microflora enzymes,namely, bacterial 7-alpha-dehydroxylase. DCA is partially absorbed fromthe intestine and involved in recirculation of bile acids after itsconjugation with taurine or glycine in the liver.

Increase of the transit time in the intestine increases DCA formation asa result of bacterial metabolism, while decrease of the transit time hasthe opposite effect.

As a result, the amount of DCA varies within a wide range—from 10% to30% of the total pool of bile acids. Recently it has been proved thatpatients with cholelithiasis have increased number of gram-positiveanaerobic bacteria, and their 7-alpha-dehydroxylase activity in thecolon is higher compared to healthy patients.

In the process, a correlation of slower transit through the intestine,higher DCA share, bile oversaturation with cholesterol and concrementformation has been found. It is assumed that DCA facilitates bilelithogenicity and concrement formation by slowing the transit timethrough the intestine, which in turn increases cholesterol absorptionand, via the positive feedback mechanism, facilitates the formation ofDCA itself. In addition, DCA can increase cholesterol secretion intobile by acting on the canalocular membrane of hepatocyte, wherecholesterol is located in sphyngomyelin domains, and also increasecholesterol crystallization in bile, destabilizing vesicles withcholesterol.

In bladder, bile BAs are mainly present in the form of binarycompounds-conjugates. As the result of BA conjugation with amino acid,glycine, glycocholic (GCA) or glycochenodesoxycholic (GCDCA) acid isformed. In BA conjugation with taurine (2-aminoethan-sulfoacidC2H703N5), the product of cycteine amino acid degradation, taurocholic(TCA) or taurodesoxycholic (TDCA) acid is formed.

BA conjugation includes stages of formation of CoA-BA esters, andlinkage of the BA molecule with glycine or taurine by means of amidebond, with the participation of lysosomal enzyme of acyltransferaze. Theratio of glycine and taurine conjugates of BA in bile, 3:1 on average,can vary depending on the composition of food and on the hormonal statusof the organism.

Thus, disturbance of bile acids metabolism is an important pathogenicfactor of development of a number of liver diseases.

Known is the method for treatment of the above liver diseases thatconsists of using UDCA preparations in the form of mono- or complextherapy (RU 2002123352 A of Mar. 27, 2004).

Also known is the method for treatment of liver diseases by using CDCApreparations in complex therapy, see Register of Medicinal Agents ofRussia. Encyclopedia of Drugs]. G. L. Vyshkovskiy Editor-in-Chief. M.,“RLS”—2006, 2005, pp. 895-896).

SUMMARY OF THE INVENTION

The present invention comprises a pharmaceutical composition for atreatment and prevention of recurrences of liver diseases in humancaused by a disturbance of lipid-cholesterol metabolism, wherein theliver diseases are selected from a group including cholelithiasis withmainly cholesterol stones, alcoholic and nonalcoholic steatohepatitis,primary biliar cirrhosis, gall bladder cholesterosis, and drug-inducedand toxic liver injury, the composition comprising a mixture of ahepatoprotector and a prebiotic combined together in effective doses ina ratio of 1:2 to 1:250 by mass of pure substances, the mixture beingtaken by a person orally, the doses leading to a restoration of liverfunctions and prevention of exacerbations of liver diseases, wherein thehepaprotector comprises ursodeoxycholic acid (UDCA) and the prebioticcomprises lactulose.

In some aspects, the hepatoprotector further comprises amino acids.

In some aspects, the hepatoprotector further comprises active componentsof milk thistle plant extracts.

In some aspects, the hepatoprotector further comprises essentialphospholipids.

In some aspects, the hepatoprotector further comprises other bile acidsor bile acid salts.

In some aspects, the bile acids or bile acid salts are selected from thegroup consisting of chenodesoxycholic acid (CDCA), desoxycholic acid(OCA), lithochloic acid (LCA), taurodesoxycholic acid (TDCA),hyodeoxycholic acid (HDCA), taurocholic acid (TCA), glycochloic acid(GCA), and combinations thereof.

In some aspects, the prebiotic further comprises fructooligosaccharides.

In some aspects, the prebiotic further comprises maltooligosaccharides.

In some aspects, the prebiotic further comprisesgalactooligosaccharides.

In some aspects, the prebiotic further comprises xylooligosaccharides.

In some aspects, the prebiotic is lactulose in a ratio of 1:2 by mass ofpure substances.

In some aspects, the hepatoprotector further comprises essentialphospholipids selected from a group consisting of phosphatidylcholin,phosphatidylethanolamine and phosphatidylinositol, in a ratio of theessential phospholipids and the prebiotic is from 1:0.1 to 1:100 by massof pure substances.

In some aspects, the hepatoprotector further comprises active componentsof milk thistle plant extracts selected from silimarin or silibin, and aratio of the active components of milk thistle plant extracts and theprebiotic is from 1:0.1 to 1:100 by mass of pure substances.

In some aspects, the composition is made in the form of tablets,granules, globules, powders or capsules, suspensions, pastes, syrups,emulsion, or gels intended for oral administration 2-3 times a day.

In some aspects, the hepatoprotector and the prebiotic are taken ineffective doses in a ratio of 1:2 to 1:50 by mass of pure substances.

Also disclosed is a method for treating and preventing recurrences ofliver diseases in humans caused by a disturbance of lipid-cholesterolmetabolism, wherein the liver diseases are selected from a groupincluding cholelithiasis with mainly cholesterol stones, alcoholic andnonalcoholic steatohepatitis, primary biliar cirrhosis, gall bladdercholesterosis, and drug-induced and toxic liver injury, the methodcomprising administering a composition orally on a patient, wherein thecomposition comprises a mixture of a hepatoprotector and a prebioticcombined together in effective doses in a ratio of 1:2 to 1:250 by massof pure substances, wherein the hepaprotector comprises ursodeoxycholicacid (UDCA) and the prebiotic comprises lactulose, and delivering theUDCA and lactulose to the patient's liver.

In some aspects, a therapeutic effect on the patient's liver developswithin 1 month.

In some aspects, a therapeutic effect on the patient's liver developswithin 3 months.

In some aspects, the administering occurs 3 times daily.

In some aspects, the UDCA:lactulose ratio is between 1:2 and 1:250.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENT

According to prior art treatment methods, the therapeutic effect onlydevelops after a prolonged period (from several months to 6-12 months)of using medications, and often requires essentially taking them forlife.

To a large extent this is due to the fact that the use of BApreparations as monotherapy cannot eliminate completely such importantpathogenic factor as intestinal dysbiosis and complex systemicdisturbances of metabolism it causes.

As drugs with hepatoprotective properties, one has long been usingsubstances with different structure and mechanisms of action; a lot ofprofessionals challenge the appropriateness of considering them truehepatoprotectors.

In particular, the class of so-called essential phospholipids is oftenadded to them.

Phospholipids, or phosphoglycerides, are highly specialized lipids; theyare important fundamental components of cell membranes and membranes ofstructural elements of cells, such as mitochondria, and can be called“essential” (irreplaceable) for growth, development and properfunctioning of all somatic cells. In addition to their role in buildingcell membranes, one can say that phospholipids are important componentsof lipoproteins, “lung surfactants” and bile. They take part inoperation of the nervous system and in membrane enzyme reactions, andplay an important role in metabolism and oxidation processes. As part oflipoproteins, phospholipids affect the level of cholesterolconcentration in blood.

Phospholipids located in trombocites participate in the blood clottingprocess, which in the end demonstrates their effect on the protectivefunction of blood and on hemodynamics in the organism of mammals andhumans. Phospholipids chemical structure, difility and presence ofcharged groups determine the uniqueness of their physiologicalproperties.

The main function of phospholipids is forming a double lipid layer incell membranes. The structure and function of cell membranes areextremely important for person's health. The feeling of general malady,disturbance of functions and various diseases can in many cases beexplained by damage to or instability of membranes. By introducingphospholipids one can affect membrane functions related to membraneproteins and correct them, at least to a certain extent, and sometimescompletely correct the disturbed function.

Essential phospholipids mainly penetrate liver cells, embed intohepatocite membranes, normalize liver functions and metabolism of lipidsand proteins, facilitate activation and protection ofphospholipid-dependent enzyme systems, improve the detoxificationfunction of the liver, restore its cell structure, improve regeneration,and inhibit formation of connective tissue in it.

The preparation reduces the level of energy consumption in the liver,converts neutral fats and cholesterol to easily metabolized forms, andstabilizes bile physical and chemical properties.

Essential phospholipids normalize intestinal digestion not only of fatsbut also, indirectly, of solid food due to restoration of the structureof liver cells, which results in normalization of bile formation andexcretion (Gurevich, K. G., Essential Phospholipids in Treatment ofLiver Diseases, see High Quality Medical Practice, 2002, No. 4, pp.108-111).

Another representative of the group of substances with hepatoprotectiveproperties is milk thistle plant extract as the main active substance insilimarin. It has hepatoprotective, regenerative and detoxificationeffect.

It neutralizes free radicals in the liver and prevents the destructionof cell structures. It specifically stimulates RNC polymerase andactivates the synthesis of structural and functional proteins andphospholipids in damaged hepatocites. It stabilizes cell membranes,prevents exit of intracellular components (transaminases) andaccelerates regeneration of liver cells. It inhibits penetration of somehepatotoxic substances (death cup mushroom poison) into the cell.

Clinical pharmacology: improves the general well-being of patients withliver diseases, reduces subjective complaints—weakness, feeling ofweight in the right hypochondrium, loss of appetite, vomiting and skinitch, and normalizes laboratory parameters—the activity oftransaminases, gamma glutamyl and alkaline phosphotase, and bilirubinlevel. In prolonged use it increases the life expectancy of patientswith liver cirrhosis.

A number of amino acids or their derivatives are also often consideredpreparations of the hepatoprotective series.

The best known of them is ademethionine, see Register of MedicinalAgents of Russia. Encyclopedia of Drugs, G. L VyshkovskiyEditor-in-Chief, M., “RLS”-2006, 2005, p. 51.

This preparation makes up for the deficiency of ademethionine andstimulates its production in the body, first of all in the liver andbrain. The S-adenosil-L-metionine (ademethionine) molecule donates themethyl group in reactions of methylation of phospholipids of cellmembranes of proteins, hormones, neuromediators, etc.(transmethylation). It is the precursor of physiological thyolcompounds-cycteine, taurine, glutathione (provides a redox mechanism ofcell detoxification), coenzyme A, etc. in transsulfatation reactions.After decarboxylation, it participates in processes of aminopropylationas the precursor of polyamines-putrescine (stimulator of cellregeneration and proliferation of hepatocites), spermidine and sperminethat are part of ribosomes structure.

It has anticholestatic effect and is effective in the intralobialversion of cholestasis (disturbance of bile synthesis and flow). Theanticholestatic effect is due to increased motility and polarization ofhepatocite membranes because of stimulation of the synthesis ofphosphatidylcholine in them. This improves the function of hepatocitesof transport systems of bile acids (BAs) associated with membranes andfacilitates BA passage into the bile excretion system. It stimulates BAdetoxification—increases the content of conjugated and sulfated BAs inhepatocites. Conjugation with taurine increases BAs solubility andelimination from hepatocite. Sulphatation makes possible elimination bykidneys and facilitates hepatocite passage through the membrane andexcretion with bile. In addition, sulphated BAs protect liver cellmembranes from toxic action of non-sulphated BAs (present in highconcentrations in hepatocites in intrahepatic cholestasis).

In patients with diffuse liver diseases (cirrhosis, hepatitis) withintrahepatic cholestasis syndrome it reduces the intensity of skin itchand changes of biochemical indices, including the level of conjugatedbilirubin, ALP activity, aminotransferases, etc.

The therapy was accompanied by disappearance of asthenic syndrome in 54%of patients and the decrease of its intensity in 46% of patients. Theantiasthenic, anticholestatic and hepatoprotective effects continued for3 months after the treatment was stopped. Their efficiency in the caseof hepatopathies caused by hepatotoxic medicinal agents (paracetomol,etc.) has been demonstrated. As a result of treatment of patients withopiate drug addiction accompanied by liver injury, regression ofclinical manifestations of abstinence, improved functional condition ofthe liver and microsomal oxidation processes, and the anti-depressanteffect were observed.

The use: intrahepatic cholestasis; liver injuries-toxic, includingalcoholic, virus and drug-induced (antibiotics; anticancer,antituberculous and antivirus preparations; tricyclic antidepressants;and oral contraceptives); cirrhotic and pre-cirrhotic conditions;encephalopathy, including encephalopathy associated with hepatic failure(alcoholic, etc.); depressive and abstinence syndromes.

In addition to directly affecting the liver tissue, ademethionine has anumber of additional pharmacological effects, such as antidepressantaction (develops during the first week and stabilizes during the secondweek of treatment). This preparation is also used empirically inosteoarthrytises; this is accompanied by reduced pain syndrome, bystimulation of the synthesis of proteoglycans, and by partialregeneration of cartilaginous issue.

However, the use of essential phospholipids in therapy of liverdiseases, as well as for other hepatoprotectors, usually does notprovide full cure, and when the preparations are cancelled, there areoften recurrences or exacerbation of liver diseases, including becauseof persistent disturbance of intestine biostenosis, which is stubborn ina number of diseases of the gastrointestinal tract organs and liver.

In recent years, the concept of unity of all processes taking place ingastrointestinal tract pathology has been getting increased recognitionin gastroenterology. Within the framework of this concept, one of themost important components of this normal state is normalization of colonmicroflora.

Microflora of the gastrointestinal tract (GIT) and the liver closelyinteract in processes of organism detoxification. Microbiota in biofilmis the first one to get in contact and subsequent metabolic reactionswith all substances entering the body with food, water or atmosphericair. Microblota converts chemical substances to non-toxic end productsor to intermediate compounds that are easily broken down in the liverand then eliminated from the body.

The body has two main detoxifying organs—the liver that performs bodyprotection by means of oxidation reactions, and digestive tractmicroflora which uses for these purposes hydrolytic reduction processes.Disturbance of interaction of these systems results in mutual functionaland structural changes in them and the body as a whole.

This is why enterohepatic circulation of various organic and inorganicsubstances can be, without exaggeration, considered among cardinalhomeostatic mechanisms. Decrease of the detoxification function of GITmicroflora in dysbiosis, caused by various pathogenes (drugs, food,stress, etc.) increases the load on enzyme systems of the liver, andunder certain conditions facilitates the appearance of metabolic andstructural changes in it.

In the case of disbalance of the digestive tract microecology, anincreased proportion of potentially pathogenic gram-negative bacteriaresults in substantial accumulation of endotoxins in the lumen of theintestine.

As endotoxins penetrate the local blood circulation system throughintestine mucosa and then enter the liver through the portal vein, theydamage hepatocites or potentiate adverse effects of other toxicants.

Ninety percent of all endotoxins are freed facultatively bygram-negative bacteria. Endotoxins damage cell membranes, disturb iontransport, cause fragmentation of nucleic acids, induce formation ofproducts of free radical oxidation, initiate apoptosis, etc. (Gracheva,N. M., et al. Chilak-Forte in Complex Treatment of Patients With AcuteIntestinal Infections and Chronic Diseases of Gastrointestinal TractWith Effects of Dysbacteriosi. Consilium medicum. 2004, No. 1, pp.31-34).

Therefore, one of the possible ways to correct disturbances in thecomplex of microbiota and liver interaction is to fight intestinedysbiosis.

Then, known is the method for normalization of intestinal microflora byoral (per os) administration of probiotics-live bacteria, species andgenera normally colonizing the colon of humans and other mammals (V. F.Dyomin et al. The Experience of Using Biophitocorrection in ChildrenWith Disbiosis, see Journal of Modern Pediatrics, 2003, No. 3, vol. 2,pp. 33-36. However, the use of probiotics in the form of monotherapydoes not produce sustainable effects because of the “foreignness” ofbacteria strains, and their fairly rapid elimination (3-5 days) afterstopping taking the medicinal agent (MP).

Other preparations used for correction of disturbances of intestinalmicrobiota-prebiotics do not have this shortcoming.

Probiotics, among other things, include a lot of oligosaccharides thatare not utilized by human organism because the intestine does not haveits own enzymes that break up such sugars. Among non-digestibleoligosaccharides are, in particular, fructooligosaccharides (FOS),maltooligosaccharides, galactooligosaccharides, inulin, lactulose andsome other oligosaccharides that can be used as prebiotics (Sheveleva,S. A., Probiotics, Prebiotics qnd Probiotic Products. State of theArt.>≦[Journal of Nutrition], 1999, No. 2, pp. 33-39; Shoaf K et al.Prebiotic galactooligosaccharides reduce adherence of EnteropathogenicEscherichia coli to tissue culture cells. Infect Immim., 2006, Sep. 18.Abstr.).

As far as their chemical structure is concerned, FOS areoligofructosaccharides, wherein β-, D-fructofuranose residues areconnected to each other by β-2, 1-glycoside bonds, and on one end of thechain they have α-glucose residue connected to fructose by α-1, 2 bond.

They can be considered derivatives of saccharose, with from 1 to 3fructofuranose residues connected to its fructose part by β-2, 1 bonds.The main components of FOS are 1-chestose (GF2), nistoze (GF3) andIF-fructofuranosylnlstose (GF4).

FOS have a pronounced prebiotic effect—they are not digested in upperGIT, suppress growth of putrid microflora, facilitate normalization ofblood pressure and the level of lipids in blood, improve adsorption ofcalcium and magnesium, increase immunity, have a beneficial effect onconstipation and purulent processes, and prevent colon cancer.

Like all prebiotics, FOS are not hydralized by GIT ferments, are notabsorbed in the small intestine and, entering the colon unchanged, are aselective substrate for growth of normal microflora.

Lactulose is a disaccharide consisting of galactose and fructose(4-O-D-galactopyranosyl-D-fructose). In vivo, lactulose in smallquantities can form when milk is heated to temperatures above 100° C.Lactulose is very soluble in water and about 1.5-2 times sweeter thanlactose.

The prebiotic effect of lactulose increases the volume of coloncontents, decreases pH, decreases ammonia content in the colon andincreases content of short-chain fatty acids, particularly propionicacid (ZDUNCZYK Z et al. Physiological effects of lactulose and inulin inthe caecum of rats. Arch Anim Nutr., 2004. Vol. 58(1), pp. 89-98).

Also known is the lactulose effect on intestinal microflora-increasingthe number of bifidobacteria with increased activity of microbialβ-galactosidases (BOUCHNIC Y et al. Prospective, randomized?Parallel-group trial to evaluate the effects of lactulose andpolyethylene glycol-4000 on colonic flora in chronic idiopathicconstipation. Aliment. Pharmacol. Ther., 2004, Vol. 19(8), pp. 889-899).

Being a prebiotic, until now lactulose has at the same time been used intherapy mainly or even exclusively as a mild and effective laxative. Thelaxative effect of lactulose is due to its prebiotic effect and iscaused by increased volume of colon contents (by about 30%) because ofthe increase in bacterial population.

For instance, known is the method for normalization of disorders ofintestinal microflora that includes the use of prebiotics, particularlyindigestible oligosaccharide (lactulose, FOS, etc.) (JP 2003-155242 ofMay 27, 2003).

In literature, there is very little information about attempts to useprebiotics for treatment of liver diseases.

According to published data (Nikitin, I. G. et al., Duphalac (Lactulose)in Treatment of Intestinal Dysbiosis in Nonalcoholic Steatohepatitis,see Clinical Prospects of Gastroenterology, Hepatology, andColoproctology], 2002, No. 1, pp. 24-29; Savelev, V. S., Lipid DistressSyndrome in Surgery and Bulletin of the Russian Military MedicalAcademy, 1999, No. 1, pp. 36-39, their use as mono-preparations is byitself has little effect when attempting to treat liver diseases becausein this case the damage to cells and, accordingly, to functions ofdamaged liver tissue is not completely eliminated.

The therapeutic effect of only using prebiotics occurs after a fairlylong period, and in this case complete restoration of disturbed lipidmetabolism and particularly of cholesterol metabolism is not achieved.Moreover, in the referenced studies lactulose was used in large doses(on the order of 30 ml of concentrated syrup per person) that are knownnot as much as prebiotic but rather laxative doses.

Usually, lactulose is used in hepatology in such large doses mainly toalleviate symptoms of liver encephalopathy, i.e. in palliativesymptomatic alleviation of condition in patients with alreadypractically incurable conditions (advanced stage cirrhoses). In thesesituations, normalization of intestinal microflora cannot provide anylong-term effect.

Therefore, in schemes of therapy of various diseases, the schemes ofcomplex application of various groups of medicinal agents, for instance,immunomodulators and antibiotics, choleretics, etc. are ever more widelyused.

In this respect, particularly known is the method for treatment ofchronic non-calculous cholecystitis with manifestations of dysbiosis dueto the use of combination of a hepatoprotector (glutargin) and aneubiotic (bifiform) (UA 70018 of Sep. 15, 2004).

However, the therapeutic result of this combination is short-term due tothe noted earlier limitations related to the use of probiotics. Alsoknown is the method for treatment of liver diseases with cholestaticsyndrome by using a combination of a phytogenic hepatoprotector (extractfrom Silibum marianum) with a probiotic strain of Lactobacillusbulgarinii and some other substances (BG 108250U of Apr. 30, 2005).

However, in this case too, the positive therapeutic effect is relativelyshort-term.

Known is the method for correction of Savelev's lipid distress syndromeusing complex therapy including a phytogenic hepatorpotector-gepabene,and metabolite probiotic chilac-forte (Petukhov, V. A., Liver FunctionAbnormality And Dysbiosis in Lipid Distress Syndrome. PM FigureUS20110312910A1-20111222-P00019 [RMZh], 2002, No. 10, Vol. 4, pp.158-160).

However, in this analogue neither the scheme of treatment using thesemeans nor their doses or ratios are specified, nor the interaction ofthe medicinal agents used is shown.

Also known is the use for treatment of liver diseases of a combinedpreparation comprising alkaline sphingomyelinase as the main actingagent, and various substances, including substances from the class ofprobiotics (Lactobacillus acidophilus, Lactobacillus brevis,Lactobacillus buchneri, Lactobacillus casei, Lactobacillus catenaforme)and UDCA, as additional means (EA 5166 of Dec. 30, 2004).

In the above analogue, the leading roles is played by sphingomyelinase(a lysosomal enzyme), which is used for prevention and/or treatment ofvarious diseases from the following group: small intestine disorder,malignant tumors, immune system disturbance, inflammations anddesquamation of the mucous membrane of the small intestine, conditionsassociated with disturbances in the synthesis of cholesterol,disturbances of absorption ability of the small intestine, and allergydiseases of the small intestine.

Herein, said pharmaceutical composition includes as additives probiotics(Lactobacillus acidophilus, Lactobacillus brevis, Lactobacilus casei,etc.), ursodeoxycholic acid—a bile acids derivative, and aprebiotic—lactulose.

But the patent description does not disclose the role of these additivesin treatment of liver diseases, and there is no scientific justificationof including them in this composition.

The use of the pharmaceutical composition for treatment of said diseasesis not without the main shortcoming—a short-term effect, caused by anexogenous probiotic strain. Herein, prebiotic components present in thecomposition are to a large degree utilized by said introduced exogenousstrain.

In addition, inclusion in the composition of a large number ofcomponents with different type of action not just does not make itpossible to evaluate their role and therapeutic effect—it does notpreclude mutual antagonism of such effects either, which increases theprobability of individual variations in the reaction to administrationof such large number of preparations.

The closest analogue of the claimed invention is the agent or componentthat improves the liver function, acts as the methyl group donor,includes a not easily assimilated oligosaccharide that comprisesgalactose, and is used as a functional nutrition product (JP 2003-155242of May 27, 2003).

Herein, the methyl group donor is selected from the group of amino acidsthat includes S-adenosylmethionine, and the galactose-containingoligosaccharide is selected from the group that includes, among others,lactulose or galactooligosaccharide.

However, in the said patent the authors do not consider prebioticeffects of the composition at all, and in the proposed agent the authorsreduced the role of the oligosaccharide component (and only supposedlyat that) to elimination of ammonia-induced hepatoencephalopaticintoxication.

By that, the authors of the closest analogue do not link the lipidmetabolism normalizing effect of compounds—the donors of methyl groupsto normalization of the condition of intestinal microbiocenosis, and theexamples provided do not mention this condition at all.

The lack of understanding of inseparable interrelation of the conditionof intestinal microbiocenosis and lipid metabolism reactions, and therole of representatives of normal microflora in breaking the viciouscircle of enteropathogenic recirculation of bile acids does not make itpossible to correctly select not just the component content of thecompositions depending on the degree of disturbance of these mostimportant components of the metabolic process, but also to control tothe necessary degree the adequacy and effectiveness of the therapy ofliver diseases that is performed.

By reducing the role of the first component of the composition just tothe function of the methyl group donor the authors unreasonably ignoreother mechanisms of disturbance of lipid metabolism, particularly ofcholesterol metabolism, that play the most important role in thedevelopment of many liver diseases.

The distinction of the claimed method from the closest analogue is asfollows:

the liver diseases that are different from the closest analogue and fortreatment of which a real pharmaceutical composition has been developedare clearly defined;

a pharmaceutical composition has been developed that comprises ahepatoprotector and a prebiotic selected strictly from a limited numberof representatives of these groups of medicinal agents; the advisabilityof simultaneous introduction of a hepatoprotector and a prebiotic in onepharmaceutical composition because of their synergistic effect on eachother is demonstrated;

the ratio of the hepatoprotector and the prebiotic in the claimedpharmaceutical composition has been developed.

The objective of the invention is to achieve substantial positiveeffects in the form of accelerating normalization of the health statusand reduction of the intensity of the disease symptoms in the case ofcomplex treatment of individual liver diseases.

Said objective is achieved by using in therapy of liver diseases acombined (complex) medicinal agent containing a mixture of ahepatoprotector and a prebiotic substance, particularly oligosaccharidesthat are not digestible in the intestine.

The technical result the invention aims to achieve is to restore liverfunctions as soon as possible and prevent disease recurrences byrestoring cholesterol metabolism and intestinal biocenosis caused bysynergistic interaction of the hepatoprotector and the prebiotic, whichalso causes prevention of the hepatoprotector side effects.

As the hepatoprotector, the Invention uses bile acids/bile acid saltsselected from the following group: GCA, GCDCA, TCA, TDCA, UDCA, CDCA,and essential phospholipids.

The presence of a hepatoprotector and a prebiotic in the claimedpharmaceutical composition ensures pronounced and sustainabletherapeutic effect due to the synergistic effect of the hepatoprotectorand the prebiotic.

Synergistic effect of a hepatoprotector, for instance, UDCA, and aprebiotic is due to the fact that UDCA normalizes intestinal digestionnot just of fats, but also of solid food, due to increased synthesis ofbile acids and UDCA itself in the liver, which facilitates normalizationof intestinal microflora, while the prebiotic by itself facilitatesmicroflora normalization due to the stimulation of growth of residentstrains, which causes manifestation of immunity-stimulating effects andother effects of normal flora. This results in improvement of digestionproperties and consequently in improved detoxification of exogenoustoxins by own microflora, which also reduces the metabolic load on theliver and facilitates normalization of metabolism of fatty acids andcholesterol, which in turn has a stabilizing effect on all body cells,including hepatocites. Thus, combining bile acids or bile acid salts inone pharmaceutical composition with oligosaccharides that are notdigestible in the small intestine and chosen from the following group:lactulose or FOS, maltooligosaccharides, galactooligosaccharides,inulin—makes it possible to restore the function of hepatocites and theliver as a whole due to normalization of intestinal biocenosis, which inturn ensures long-term stabilization of the achieved therapeutic result.

It is well known that in liver diseases and cholelithiasis with mainlycholesterol stones, one observes higher content of toxic products inpatient's blood that enter blood from the colon, especially of ammonia

a nitro compound that forms in the process of bacterial decomposition ofprotein by proteolytic microflora in the colon, which increases thetoxic load on the liver.

Thus, restoration of microflora in the intestine indirectly facilitatesthe increase decrease [sic] of the toxic load on the liver and theincrease of UDCA in the liver, which in turn facilitates restoration ofbile composition, particularly the increase of bile acids due todecreased synthesis of cholesterol by the liver, and this in turnsprevents recurrences of the disease, particularly of cholelithiasis withmainly cholesterol stones.

Taking the above into account, active components in the presentedpharmaceutical composition mutually amplify the therapeutic propertieseach of them has.

The claimed pharmaceutical composition can include as the active agentvarious substances with hepatoprotective properties, particularly ahepatoprotector selected from the following groups: amino acids or theirderivatives, active hepatoprotective substances from milk thistle plantextracts (silimarin, silibinin), or essential phospholipids, or bileacids/bile acid salts selected from the following group: CA, CDCA, DCA,UDCA, HDCA, TUDCA, TCA, and GCA in the unit dose from 50 mg to 500 mg,and a prebiotic selected from the group of oligosaccharidesnon-digestible in a human intestine, such as lactulose, orfructooligosaccharide (FOS), or maltooligosaccharides, orgalactooligosaccharides, or inulin, taken each in the effectivetherapeutic prebiotic dose, galactooligosaccharides,maltooligosaccharides or xylooligosaccharides, with the hepatoprotectorto prebiotic ratio from 1:2 to 1:250.

The claimed pharmaceutical composition can be made in the form oftablets (coated or uncoated), or granules, or globules, or powder, orcapsules, or suspensions, or emulsions, or gels.

In doing so, the claimed pharmaceutical composition can additionallycomprise additives that are generally accepted in the pharmaceuticalindustry, such as microcrystalline cellulose or lactose, or corn starch,or potato starch, or hydroxypropylmethylcellulose, orcarboxymethylcellulose, or oxypropylmethylcellulose, oroxypropylcellulose, or their pharmaceutically acceptable salts, orludipress, or calcium stearate, or magnesium stearate, or polysorbate,or polyvinyl pyrrolidone, or polyethylene glycol, or talcum, or titaniumdioxide, or silicon dioxide.

The claimed composition is prepared by mixing the components comprisingit, both active (the hepatoprotector selected from the following group:essential phospholipids or bile acids/bile acid salts, and theprebiotic) and additives selected from the following group:microcrystalline cellulose, or lactose, or corn starch, or potatostarch, or hydroxypropylmethylcellulose, or carboxymethylcellulose, oroxypropylmethylcellulose, or oxypropylcellulose, or theirpharmaceutically acceptable salts, or ludipress, or calcium stearate, ormagnesium stearate, or polysorbate, or polyvinyl pyrrolidone, orpolyethylene glycol, or talcum, or titanium dioxide, or silicon dioxide.

The claimed pharmaceutical composition is administered orally, washeddown with large amount of water, for a period from 1.5 to 3 months.

The claimed pharmaceutical composition can be used for treatment ofpatients with liver diseases selected from the following group:cholelithiasis with mainly cholesterol stones, alcoholic andnon-alcoholic steatohepatitis, primary biliary cirrhosis, gall bladdercholesterosis, and drug-induced and toxic liver injury, and it makes itpossible to achieve long-term remission of the disease in a relativelyshort time period (from 6 to 12 weeks).

Herein, the therapeutic effectiveness is from 89% to 95%.

The claimed pharmaceutical composition does not have contraindicationsand can be used for treatment of patients with liver diseases listedabove, including against the background of severe concomitant diseases(except advanced stages of liver cirrhosis, malignant tumors ofgastrointestinal tract or other organs), regardless of patient's age.

The claimed composition does not have significant side effects, becausethe active components in the pharmaceutical composition are used insmall and medium unit therapeutic doses and during a fairly short timeperiod.

The proposed versions of the composition are characterized by low costand are therefore affordable for all categories of patients.

Patient treatment is performed outpatient and does not require thepatients to keep bed rest or semi-strict bed rest regimen, which makesit possible for patients to lead normal life.

The proposed pharmaceutical composition can be successfully used notjust for treatment but also for prevention of exacerbations of liverdiseases selected from the following group: cholelithiasis with mainlycholesterol stones, alcoholic and non-alcoholic steatohepatitis, primarybiliary cirrhosis, gall bladder cholesterosis, and drug-induced andtoxic liver injury, due to restoration of the structure and function ofboth hepatocites and function of the liver as a whole caused bynormalization of intestinal microflora. The effect of the proposedpharmaceutical composition in a certain dose on the body stipulatesgradual and increased enhancement of the therapeutic effect andconnecting new levels of homeostasis regulation—the sub-cell,inter-cell, tissue, organ, system and body level, due to restoration oflipid metabolism, particularly cholesterol metabolism because ofnormalization of intestinal microbiocenosis.

During treatment, which is performed outpatient, it is recommended thatthe patient keep at least three meals a day regimen during the entiretreatment period; it is not recommended to take alcohol, fatty and spicyfood and other medicinal agents; fasting and hard physical work areprohibited.

The final diagnosis is determined based on additional types ofexaminations (USE of the liver or radiology of bile passages) andlaboratory blood tests (biochemistry: cholesterol and its fractions,biliburin and its fractions, alkaline phosphotase, GPT, AST, ESR, etc.).

We examined 60 patients. In all patients, clinical symptoms of liverinjury with various degree of manifestation had been found: ochrodermiaof skin and sclera, skin itch, sense of discomfort or feeling of weightin the right hypochondrium, dyspeptic effects-nausea, anorexia,vomiting, weakness, atony, change of the color of urine (darker) andstool (loosening or diarrhea).

All patients had been treated earlier either outpatient or inpatient,using various medicinal agents. Most patients (37 out of 60) hadconcomitant diseases: chronic gastroduodenitis and cardiovascular system(CVS) diseases: hypertensive disease (HD), ischemic heart disease (IHD);lung diseases: pneumosclerosis, bronchial asthma (BrA), etc. Besides, in85% of patients concomitant disturbances of the state of colonbiocenosis were identified. For all patients in experimental groups(50), treatment using the claimed pharmaceutical composition wasperformed in the dosing regimen that had been developed: 3 times a dayat meal time for 1.5-3.0 months.

By the end of the 2nd week, the detoxification and synthetic function ofthe liver had been restored in all 50 patients regardless of thecharacter of liver injury, against the background of restoration ofintestinal biocenosis.

All patients noted relief of discomfort in the right hypochondrium andimproved general health as early as by the 5th day. By the start of the2nd week the disappearance of dyspeptic disorders, restoration ofappetite, normalization of urine and stool, disappearance of skin itchand restoration of the original skin color against the background ofsignificant improvement in general condition and mood was observedsubjectively in all patients; with this, the simultaneous relief ordisappearance of concomitant pathology symptoms were noticed. Inbiochemical blood tests by the end of the 3rd week, normalization of allbiochemical indexes, including those that characterize liver operationand lipid metabolism was noted.

Examples of Embodiment of the Method 1. Patient I., Male, 46 Years Old

On admission, complaints of aching pain in the right hypochondrium,radiating to the right shoulder, appearing 3-4 hours after consumingfatty food, or after a lavish meal, or after physical exertion; ofgeneral weakness, anorexia, nausea, periodic vomiting, feeling ofbitterness in the mouth, stool loosening, sometimes diarrhea, skin itch,change of the color of urine (darker) and stool (lighter).

From the anamnesis: cholelithiasis for 10 years. Has not been operated.Has been treated outpatient, without much effect. Worsening afterphysical exertion.

Objectively: supernutrition, weight 75 kg, height 167 cm, skin pale,with traces of scratching on the back and abdomen. Icteric sclera.

Abdomen soft and painful at the Kehr point. Kehr's, Mussy's and Murphy'ssymptoms positive. Liver at the edge of the coastal arch. Vesicularbreathing in the lungs. Breathing rate 18 a minute.

Cardiac border within the age norm. Sounds moderately deadened, rhythmregular, heart rate 78 a minute, BP 140/85 mm Hg.

Pasternatsky's syndrome negative on both sides. Provisional diagnosis:Chronic cholecystitis in the exacerbation phase.

Examination:

Complete blood count: Hb 123 g/l; erythrocytes (ER) 4.11×1012/l; colorindex (CI) 0.89; leukocytes 4.0×109/l; stab (S) leukocytes (L) 2%;segmental leukocytes (S) 46%; eosinophils (E) 5%; lymphocytes (L) 45%;monocytes (M) 2%; ESR 40 mm/h. Complete urinalysis: relative density1016; no protein or glucose detected; leukocytes 0-1-3 in field of view;erythrocytes 0 in field of view; urine amilase 16.2 mgs/l.

Coprogram: muscle fibers without striation—a little; fattyacids—moderate amount; undigested phytogenic fiber—a lot; starch;isolated cells.

Feces on dysbacteriosis: Reduction of the number of bifidobacteria andlactobacilli, respectively: 105/g and 106/g, due to increase of Candidafungi. Blood biochemistry: bilirubin and its fractions: total bilirubin(TB)—22.8 μmol/l (N—3.4-20.5 μmol/l); conjugated bilirubin (CB)—3.8μmol/l (N—0.85-3.4 μmol/l), non-conjugated bilirubin (NCB)—11.7 μmol/l(N—2.56-10.3 μmol/l); thymol test (TT)—12.1 units (N—4 units), ACT—79units (N—60 units) GPT—72 units (N—50 units), thymol test (TT)—1.7 units(N—4 units), alkaline phosphatase (ALP)—362 units (N—up to 295 units),CCC 15.3; sugar 3.5 mmol/l (N—4.4-6.6 mmol/l);

cholesterol and its fractions: total cholesterol (TC)—5.5 mmol/l(N—3.65-5.2 mmol/l), high-density lipids cholesterol (HDLC)—0.8 mmol/l(N—0.9-1.9 mmol/l), low-density lipids cholesterol (LDLC)—3.2 mmol/l(N—1.91-2.6 mmol/l), cholesterol atherogenic ratio (CAR) 3.5 c.u. (N—upto 3 c.u.), CCC 15.3 (N—up to 12);protein fractions: total protein 67 g/l (N 65-85 g/l); albumins 34 g/i(N—36-50 g/l);antinuclear antibodies: AMA titer 1:10;coagulogram: PTR 24 s—79%; thrombine clotting time 35 s; free heparin 12s; fibrinogen 2.2 g/l; fibrinolytic activity >240 min.

Coprogram: dysbacteriosis due to reduction of lactobacilli andbifidobacteria: lactobacilli (105) (N>=107/g), bifidobacteria (107)(N>=109).

Radiography of the liver and bile passages—indirect signs of calculouscholecystitis, stones do not contrast. Recommended: USE of the liver andgallbladder.

USE of the liver—chronic cholesistisis, cholesterol stones: 0.9, 1.2,1.5, 1.3 mm, edges even.

EKG—sinus rhythm, signs of moderate left ventricular hypertrophy. BP150/85 mm Hg; heart rate 74 a minute.

Final diagnosis: Lipid metabolism disorder, hypercholesterinemia.Chronic calculous cholecystitis (cholesterol stones) in the exacerbationphase.

Treatment: the use of the claimed composition, wherein the active agentsare a hepatoprotector—UDCA and lactulose in the ratio of 1:2 (unit doseof UDCA is 325 mg), orally 3 times a day at meal time for 1.5 monthsagainst the background of diet No. 5.

Follow-up examination in 1.5 months:

In USE: isolated stones, 1 and 2 mm in size.

According to laboratory examination, no pathology detected.

Recommended: continue the therapy for up to 3 months.

After 3 months: in USE—signs of chronic cholecystitis, no concrements.

Conclusion: chronic cholecystitis in the remission phase.

2. Patient B., Male, 45 Years Old

On admission, complaints of anorexia, weakness, nausea, periodicvomiting, aching pain in the right hypochondrium after a large or fattymeal. In anamnesis: chronic alcoholism. Primary biliary cirrhosis. Hadbeen treated irregularly.

Objectively: subnutrition. Dry skin, hot to the touch. Light yellowskin, icteric sclera.

Above the lungs—pulmonary bandbox sound. Diminished breath sounds,diffused dry rales over the entire lung surface. Breathing rate 20 aminute.

Cardiac border expanded 1.0 cm to the left. Deadened sounds. Regularrhythm, second sound accent above the aorta.

Soft abdomen, the right edge of the liver protrudes 2.0 cm from underthe coastal arch, the edge is solid. Spleen not enlarged.

Pasternatsky's syndrome doubtful.

Provisional diagnosis: primary biliary cirrhosis?

Examination:

Complete blood count: Hb 117 g/l; erythrocytes (Er) 3.5×10/l; colorindex (CI) 0.9; leukocytes 4.0×10%; stab (S)—17%, leukocytes (L) 6%;segmental leukocytes (C) 36%; eosinophils (E) 5%; lymphocytes (L) 35%;monocytes (M) 1%; ESR 40 mm/h.

Complete urinalysis: relative density 1012; no protein or glucosedetected; leukocytes 0-2-3 in field of view; erythrocytes 0-2 in fieldof view; urine amilase 14.7 mgs/l.

Coprogram: muscle fibers without striation—a little; fattyacids—moderate amount; phytogenic fiber.

Blood Biochemistry:

bilirubin and its fractions: total bilirubin (TB)—28.4 μmol/l(N—3.4-20.5 μmol/l); conjugated bilirubin (CB)—4.8 μmol/l (N—0.5-3.4μmol/l), non-conjugated bilirubin (NCB)—15.0 μmol/l (N—2.56-10.3μmol/l);

thymol test (TT)—16.1 units (N—4 units), ACT—90 units (N—60 units)GPT—74 units (N—50 units), alkaline phosphatase (ALP)—700 units (N—up to295 units), sugar 6.6 mmol/l (N—44-6.6 mmol/l);cholesterol and its fractions: total cholesterol (TC)—5.9 mmol/l(N—3.65-5.2 mmol/l), high-density lipids cholesterol (HDLC)—10.8 mmol/l(N—0.9-1.9 mmol/l), low-density lipids cholesterol (LDLC)—3.6 mmol/l(N—1.91-2.6 mmol/), cholesterol atherogenic ratio (CAR) 3.9 c.u. (N—upto 3 c.u.), CCC 16.3 (N—up to 12); (norm up to 50); protein fractions:total protein 63 g/l (N 65-85 g/W); albumins 34 g/l (N—36-50 g/l;antibodies: AMA in titer 1:45;coagulogram: PTR 24 s—79%; thrombine clotting time 31 s; free heparin 11s; fibrinogen 2.0 g/l; fibrinolytic activity>221 min.

Feces on dysbacteriosis: reduction of lactobacilli and bifidobacteria:lactobacilli (104), bifidobacteria (106).

Radiography of the liver and bile passages—the liver size increased by2.5 cm due to the right lobe, distinct edges, indirect signs of biliarcirrhosis. Recommended: USE of the liver and gallbladder.

USE of the liver—signs of slight fatty infiltration of the liver andgallbladder cholesterosis. Pancreas not enlarged. Intra- andextrahepatic bile ducts not dilated. No signs of portal hypertensiondetected.

No portal hypertension detected [sic].

Liver biopsy: Dilated portal tracts infiltrated by lymphocytes, plasmacells, macrophages and eosinocytes. Among the cells of portal tractinfiltrates there are formed lymphoid follicles. Infiltrates aredetected in walls of some intralobular bile ducts. Here and there, theintegrity of the bile ducts basic membrane is violated. Near damagedbile ducts there are granulomas built of epithelioid and giganticmulinucleate cells.

Conclusion: biliary cirrhosis.

EKG—sinus rhythm, signs of moderate left ventricular hypertrophy. BP150/85 Hg; heart rate 76 a minute.

Final diagnosis: lipid metabolism disorder, hypercholesterinemia.Primary biliar cirrhosis.

Treatment: the use of the claimed composition, wherein the active agentsare a hepatoprotector—CDCA, and FOS in the ratio of 1:250 (unit dose ofCDCA is 250 mg), orally 3 times a day at meal time for 1.5 monthsagainst the background of diet No. 5.

Follow-up examination in 1.5 months:

In USE: positive dynamics—reduction of fatty infiltration of the liver.In blood tests: reduction of hypercholesterinemia OX—4.6 mmol/l,HDLC—1.2 mmol/l, CCC 13.2; TB—21.5 μmol/l, AMA 1:30; total protein—72g/l, albumins—34%, blood sugar 5.3 mmol/l, alkaline phisohatase-301units, ACT—70 units, GPT—62 units, TT—8.1 units.

Feces test on dysbacteriosis: lactobacilli 106, bifidobacteria-107.

Recommended: continue the therapy for up to 3 months.

After 3 months—AMA 1:15, bifidobacteria 109/g, lactobacilli 107/g.

Conclusion: first degree primary biliary cirrhosis (significant positivedynamics).

3. Patient F., Female, 50 Years Old

On admission: complaints of apparent weakness, feeling of weight andaching pain in the upper right abdomen area that appear for no apparentreason, anorexia, nausea, periodic vomiting, feeling of bitterness inthe mouth, stool softening, sometimes diarrhea, skin itch, changed colorof urine (lighter) and stool (lighter).

From anamnesis: has had type 2 diabetes for 15 years, has been takingbukarban, a hypoglycemic agent. Has been under medical supervision by anendocrinologist and the district physician. Has been undergoing regularinpatient treatment at the endocrinology department, but without mucheffect. Associates the latest aggravation with virus infection(protracted course; complication: acute bronchitis, was takingantibacterial agents—cephalosporins).

Objectively: supernutrition-third degree obesity, body mass index (BMI)34, pale skin, with traces of scratches on the abdomen and inner thighs.Icteric sclera.

Abdomen greatly increased in size; soft, painful at Kehr's point. Theliver protrudes 2.5 cm from under the coastal arch, the edge is solid.Spleen not enlarged.

Diminished vesicular breathing in the lungs (because of fatty tissue).Breathing rate 22 a minute.

Heart borders extended 1.0 cm to the right, 1.5 cm to left. Deadenedsounds, regular rhythm, soft systolic murmur over the apex of the heart.Heart rate 76 a minute, BP 160/85 mm Hg.

Pasternatsky's symptom doubtful on both sides.

Provisional diagnosis: fatty hepatosis (?), type 2 diabetes, thirddegree obesity.

Examination:

Complete blood count: Hb 121 g/l; erythrocytes (Er)—4.15×1012/l; colorindex (CI)—0.89; leukocytes—3.8×10/l; stab (S) leukocytes (L)—7%;segmental leukocytes (S)—40%; eosinophils (E)—5%; lymphocytes (L) 45%;monocytes (M) 3%; ESR 39 mm/h. Blood glucose—6.8 mmol/l.

Complete urinalysis: relative density 1016; protein—traces; leukocytes3-5 in field of view; erythrocytes 0 in field of view; urine amylase16.2 mgc/l. Coprogram: muscle fibers without striation—a little; fattyacids—moderate amount; undigested phytogenic fiber—a lot; starch;isolated cells.

Feces on dysbacteriosis: Reduction of the number of bifidobacteria andlactobacilli, 105/g and 106/g (respectively) due to increased Candidafungi.

Blood Biochemistry:

bilirubin and its fractions: total bilirubin (TB)—27.0 μmol/l (N3.4-20.5 μmol/l); conjugated bilirubin (CB)—3.6 μmol/l (N—0.85-3.4μmol/l, non-conjugated bilirubin (NCB)—11.2 μmol/l (N—2.56-10.3 μmol/l);thymol test (TT)—8.0 units (N—4 units), ACT—69 units (N—60 units) GPT—76units (N—50 units), thymol test (TT)—1.7 units (N—4 units), alkalinephosphatase (ALP)—346 units (N—up to 295 units), CCC 15.3; sugar 6.9mmol/l, (N—4.4-6.6 mmol/l);cholesterol and its fractions: total cholesterol (TC)—5.9 mmol/l(N—3.65-5.2 mmol/l), high-density lipids cholesterol (HDLC)—0.8 mmol/l(N—0.9-1.9 mmol/l), low-density lipids cholesterol (LDLC)—3.6 mmol/l(N—1.91-2.6 mmol/l), cholesterol atherogenic ratio (CAR) 3.8 c.u. (N—upto 3 c.u.), CCC 16.3 (N—up to 12);triglycerides: 1.94 mmol/l (N—0.45-1.82 mmol/l)protein fractions: total protein 67 g/l (N—65-85 g/l); albumins 38 g/l(N—36-50 g/l).coagulogram: PTR 24 s—79%; thrombine clotting time 35 s; free heparin 12s; fibrinogen 2.2 g/l; fibrinolytic activity>240 min.

Coprogram: dysbacteriosis due to reduction of lactobacilli andbifidobacteria—lactobacilli (104) (N>=107/g), bifidobacteria (106)(N>=109/g).

Radiography of the Liver and Bile Passages:

uniform enlargement of the liver, no concrements.

Recommended: USE of the liver and bile passages.

USE of the liver—second degree hepatomegalia. Pancreas not enlarged.

Intra- and extrahepatic bile ducts not dilated. Signs of portalhypertension.

Esophagogastroduodenoscopy: esophageal veins dilatation B/3.

EKG—sinus rhythm, signs of moderate left ventricular hypertrophy,incomplete right bundle-branch block, BP 160/85 mm Hg;

Heart rate 74 a minute.

Final diagnosis: lipid metabolism disorder, triglyceridemia. Fattyhepatosis. Type 2 diabetes, third degree obesity.

Treatment: the use of the claimed composition, wherein the active agentsare a hepatoprotector—essential phospholipids, and FOS in the ratio of1:50 (the unit dose of essential phospholipids is 50 mg), orally 3 timesa day at meal time for 1.5 months against the background of diet No. 5.

Follow-up examination in 1.5 months:

The patient notes reduction of skin itch, increased activity; itchpractically does not bother her, notes a 5 kg weight reduction. In USE:reduction of the liver size noted. Conclusion: fatty hepatosis, firstdegree hepatomegaly.

According to laboratory examination, reduction of triglycerides—1.82mmol/l and total cholesterol—5.0 mmol/l, blood sugar has normalized-4.9mmol/l,

In feces analyses, significant increase of lactobacilli andbifidobacteria—107/g and 109/g (respectively), is noted; no Candidafungi detected.

Recommended: continue the therapy for up to 3 months.

After 3 months, in USE—signs of moderate hepatomegaly. Blood tests—nopeculiarities. Intestinal microflora has been restored: lactobacilli107/g, bifidobacteria 109/g.

Conclusion: fatty hepatosis, signs of moderate hepatomegaly. Type 2diabetes; compensation; third degree obesity.

Example 4 Testing Acute Toxicity of the Compositions

The composition of UDCA and lactulose in the ratio of 1:2 (group 1),UDCA and FOS in the ratio of 1:50 (group 2), essential phospholipids(lecithin) and galactooligosaccharides in the ratio of 1:30 (group 3),ademethioninea and lactulose in the ratio of 1:50 (group 4) wasadministered orally to outbred white mice with body mass of 15-20 g.

The control group (group 6) was administered the equal amount of starchsuspension. The animals were monitored for 4 days, and their generalcondition (appearance, mobility (activity), the regularity of food andwater intake, the appearance and character of excrements) was recorded.

Maximum Survival Excrement Amount of Rate Ap- Mo- Appearance MixtureAdministered (Alive/ pear- til- And (Group) Preparation (g) Died) anceDiet ity Character 1 2 6/0 N N N N 2 2 6/0 N N N N 3 2 6/0 N N N N 4 26/0 N N N N 5 2 6/0 N N N N 6 2 6/0 N N N N

Based on the results of examination, no evidence of acute toxicity ofeach composition under the experimental conditions-all mixtures pertainto the class of low toxicity substances (LD50 values exceed 100 g/kg ofbody mass).

Thus, the claimed pharmaceutical composition that includes as activeagents a hepatoprotector and a prebiotic selected from non-digestible inthe intestine oligosaccharides can be recommended for use under clinicalconditions for treatment and prevention of liver diseases selected fromthe following group: cholelithiasis, fatty hepatosis and nonalcoholicsteatohepatitis, primary biliary cirrhosis, gall bladder cholesterosis,and drug-induced and toxic liver injury.

What is claimed is:
 1. A pharmaceutical composition for a treatment andprevention of recurrences of liver diseases in human caused by adisturbance of lipid-cholesterol metabolism, wherein the liver diseasesare selected from a group including cholelithiasis with mainlycholesterol stones, alcoholic and nonalcoholic steatohepatitis, primarybiliar cirrhosis, gall bladder cholesterosis, and drug-induced and toxicliver injury, the composition, comprising: a mixture of ahepatoprotector and a prebiotic combined together in effective doses ina ratio of 1:2 to 1:250 by mass of pure substances, the mixture beingtaken by a person orally, the doses leading to a restoration of liverfunctions and prevention of exacerbations of liver diseases, wherein thehepaprotector comprises ursodeoxycholic acid (UDCA) and the prebioticcomprises lactulose.
 2. The pharmaceutical composition according toclaim 21, wherein the hepatoprotector further comprises amino acids. 3.The pharmaceutical composition according to claim 21, wherein thehepatoprotector further comprises active components of milk thistleplant extracts.
 4. The pharmaceutical composition according to claim 21,wherein the hepatoprotector further comprises essential phospholipids.5. The pharmaceutical composition according to claim 21, wherein thehepatoprotector further comprises other bile acids or bile acid salts.6. The pharmaceutical composition according to claim 25, wherein thebile acids or bile acid salts are selected from the group consisting ofchenodesoxycholic acid (CDCA), desoxycholic acid (DCA), lithochloic acid(LCA), taurodesoxycholic acid (TDCA), hyodeoxycholic acid (HDCA),taurocholic acid (TCA), glycochloic acid (GCA), and combinationsthereof.
 7. The pharmaceutical composition according to claim 21,wherein the prebiotic further comprises fructooligosaccharides.
 8. Thepharmaceutical composition according to claim 21, wherein the prebioticfurther comprises maltooligosaccharides.
 9. The pharmaceuticalcomposition according to claim 21, wherein the prebiotic furthercomprises galactooligosaccharides.
 10. The pharmaceutical compositionaccording to claim 21, wherein the prebiotic further comprisesxylooligosaccharides.
 11. The pharmaceutical composition according toclaim 21, wherein the prebiotic is lactulose in a ratio of 1:2 by massof pure substances.
 12. The pharmaceutical composition according toclaim 21, wherein the hepatoprotector further comprises essentialphospholipids selected from a group consisting of phosphatidylcholin,phosphatidylethanolamine and phosphatidylinositol, in a ratio of theessential phospholipids and the prebiotic is from 1:0.1 to 1:100 by massof pure substances.
 13. The pharmaceutical composition according toclaim 21, wherein the hepatoprotector further comprises activecomponents of milk thistle plant extracts selected from silimarin orsilibin, and a ratio of the active components of milk thistle plantextracts and the prebiotic is from 1:0.1 to 1:100 by mass of puresubstances.
 14. The pharmaceutical composition according to claim 21made in the form of tablets, granules, globules, powders or capsules,suspensions, pastes, syrups, emulsion, or gels intended for oraladministration 2-3 times a day.
 15. The pharmaceutical compositionaccording to claim 21, wherein the hepatoprotector and the prebiotic aretaken in effective doses in a ratio of 1:2 to 1:50 by mass of puresubstances.
 16. A method for treating and preventing recurrences ofliver diseases in humans caused by a disturbance of lipid-cholesterolmetabolism, wherein the liver diseases are selected from a groupincluding cholelithiasis with mainly cholesterol stones, alcoholic andnonalcoholic steatohepatitis, primary biliar cirrhosis, gall bladdercholesterosis, and drug-induced and toxic liver injury, the methodcomprising: administering a composition orally on a patient, wherein thecomposition comprises a mixture of a hepatoprotector and a prebioticcombined together in effective doses in a ratio of 1:2 to 1:250 by massof pure substances, wherein the hepaprotector comprises ursodeoxycholicacid (UDCA) and the prebiotic comprises lactulose, and delivering theUDCA and lactulose to the patient's liver.
 17. The method of claim 16,wherein a therapeutic effect on the patient's liver develops within 1month.
 18. The method of claim 16, wherein a therapeutic effect on thepatient's liver develops within 3 months.
 19. The method of claim 16,wherein the administering occurs 3 times daily.
 20. The method of claim16, wherein the UDCA:lactulose ratio is between 1:2 and 1:250.